miR-325-3p靶向CLDN1基因调控胃癌上皮间质转化和侵袭转移

miR-325-3p Regulates Epithelial-mesenchymal Transition,Invasion and Metastasis of Gastric Cancer via Targeting CLDN1 Gene

目的探讨miR-325-3p靶向CLDN1基因对胃癌上皮间质转化和侵袭转移的影响。方法选取人胃黏膜上皮细胞株GES-1以及人胃癌细胞株HGC27、SGC-7901、MKN-45和MGC-803,并检测细胞中miR-325-3p和CLDN1的表达。双荧光素酶报告实验验证miR-325-3p和CLDN1的靶向关系,干预胃癌细胞中miR-325-3p和CLDN1的表达,qRT-PCR和Western blot检测细胞中N-cadherin、Vimentin和MMP2的表达,CCK-8检测细胞增殖活力,Transwell和流式细胞仪分别检测细胞侵袭和凋亡能力。结果相对于GES-1细胞,MGC-803细胞中miR-325-3p表达降低而CLDN1表达增高(均P<0.05),双荧光素酶报告实验证实CLDN1为miR-325-3p的靶基因。过表达miR-325-3p能够抑制胃癌细胞的增殖、侵袭和上皮间质转化,促进胃癌细胞凋亡,抑制miR-325-3p则能够促进胃癌细胞的增殖、侵袭和上皮间质转化,抑制胃癌细胞凋亡(均P<0.05)。而过表达CLDN1则能够逆转miR-325-3p过表达对胃癌细胞生物学行为的影响。结论miR-325-3p能够靶向抑制CLDN1进而抑制胃癌细胞的侵袭转移和上皮间质转化,促进胃癌细胞凋亡,miR-325-3p有望成为治疗胃癌的新靶点。

Objective To investigate the effects of miR-325-3p on the EMT,invasion and metastasis of gastric cancer cells by targeting CLDN1 gene.Methods We selected human gastric epithelial cell lines GES-1 and gastric cancer cell lines HGC27,SGC-7901,MKN-45 and MGC-803,and detected the expression of miR-325-3p and CLDN1.The targeting relation between miR-325-3p and CLDN1 were verified by dual luciferase report experiments,and the expression of miR-325-3p and CLDN1 in gastric cancer cells were intervened.qRT-PCR and Western blot were adopted to detect N-cadherin,vimentin and MMP2 expression in cells.CCK-8 assay,Transwell assay,flow cytometry were utilized to detect cell proliferation activity,invasion and apoptosis,respectively.Results Compared with GES-1 cells,miR-325-3p expression was decreased while CLDN1 expression was increased in MGC-803 cells(P<0.05).CLDN1 was a target gene of miR-325-3p.Overexpression of miR-325-3p could inhibit the proliferation,invasion and EMT of gastric cancer cells,while promote the apoptosis of gastric cancer cells.However,the inhibition of miR-325-3p had the opposite effect(P<0.05).The overexpression of CLDN1 could reverse the effect of miR-325-3p overexpression on the biological behavior of gastric cancer cells.Conclusion miR-325-3p can suppress CLDN1,inhibit the invasion,metastasis and EMT while promote the apoptosis of gastric cancer cells.miR-325-3p is expected to be a new target in gastric cancer treatment.